Cholangiocarcinoma (CCA) is a highly metastatic tumor with poor responses to traditional chemotherapeuticagents. We have focused on new drugs which can exert effects either alone or in combination with available agentsfor a better treatment of CCA and recently reported efficacy of cepharanthine (CEP), a natural biscoclaurinealkaloid extract with anti-proliferative activity against human CCA cell lines. CEP treatment effectivelysuppressed tumor growth in CCA-inoculated mice without serious side-effects and also increased cell apoptosisin primary histocultures of CCA patient tissues, suggesting therapeutic potential of CEP against human CCA.In the present study, we further showed anti-metastatic effects of CEP on migration and invasion of human CCAcell lines. CEP at 10 μg/mL effectively suppressed migration activity of KKU-M213 and KKU-M214 as shown bywound healing and Boyden chamber assays. Similar suppressive effects were observed in invasion assays. Themolecular mechanism underlying CEP actions on cell motility and invasion could be shown to involve suppressionof ICAM-1 and MMP-2. Our results suggest CEP as a potential therapeutic agent for treating metastatic CCA.